Difference between revisions of "Part:BBa K2992044"

Revision as of 12:15, 1 October 2019

FAST reporter construct with PntnH 5-UTR+RBS.

Usage and Biology

This parts entry represents a FAST reporter construct under the regulatory control of PntnH for the non-toxic non-haemagglutinin ntnH gene of C. botulinum. The construct comprises the strong clostridial promoter PntnH BBa_K2992001 and its associated 5’-UTR containing the RBS BBa_K2992015 driving the expression of the fluorescent reporter gene FAST BBa_K2992000. Transcirptional terminator occurs through the activity of the strong clostridial terminator TFad BBa_K2284012. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from Halorhodospira halophila and has been codon optimised for fluorescence studies in the genus Clostridium. In our project we couple FAST with the natural promoters of the BotR regulon thus linking reporter fluorescence with botulinum neurotoxin production. In doing so, we hoped to generate our surrogate host strain as a model for predicting neurotoxin production in foodstuffs following food manufacturing processes.

Characterisation

Data incoming.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 330

References

Heap modular Street et al 2019. Update

@@ Line 4: / Line 4: @@
 ===Usage and Biology===
-This parts entry represents a FAST reporter construct under the regulatory control of P<i>ntnH</i> for the non-toxic non-haemagglutinin <i>ntnH</i> gene of <i>C. botulinum</i>. The construct comprises the strong clostridial promoter P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] and its associated 5’-UTR containing the RBS [https://parts.igem.org/Part:BBa_K2992015 BBa_K2992015] driving the expression of the fluorescent reporter gene FAST [https://parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcriptional terminator occurs through the activity of the strong clostridial terminator T<i>Fad</i> [https://parts.igem.org/Part:BBa_K2284012 BBa_K2284012]. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from Halorhodospira halophila and has been codon optimised for fluorescence studies in the genus Clostridium. In our project we couple FAST with the natural promoters of the BotR regulon thus linking reporter fluorescence with botulinum neurotoxin production. In doing so, we hoped to generate our surrogate host strain as a model for predicting neurotoxin production in foodstuffs following food manufacturing processes.   <br><br>
+This parts entry represents a FAST reporter construct under the regulatory control of P<i>ntnH</i> for the non-toxic non-haemagglutinin <i>ntnH</i> gene of <i>C. botulinum</i>. The construct comprises the strong clostridial promoter P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] and its associated 5’-UTR containing the RBS [https://parts.igem.org/Part:BBa_K2992015 BBa_K2992015] driving the expression of the fluorescent reporter gene FAST [https://parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcirptional terminator occurs through the activity of the strong clostridial terminator T<i>Fad</i> [https://parts.igem.org/Part:BBa_K2284012 BBa_K2284012]. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from Halorhodospira halophila and has been codon optimised for fluorescence studies in the genus <i>Clostridium</i>. In our project we couple FAST with the natural promoters of the BotR regulon thus linking reporter fluorescence with botulinum neurotoxin production. In doing so, we hoped to generate our surrogate host strain as a model for predicting neurotoxin production in foodstuffs following food manufacturing processes.   <br><br> <br><br>
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 ===Characterisation===
 Data incoming.